This experiment was conducted to establish an in vitro regeneration system for four genotypes, 'Guilio', 'Cesare', 'Augustus', 'Silla' of Cichorium intybus L. var. sativus, using leaf and petiole segments as explants. The basal media consisted of Murashige and Skoog's macro elements, Morel and Wetmore's vitamins, Gamborg's micro nutrients, and 3.5 mM morpholinoethane-sulfonic acid (MES). The medium to which was added BAP 1.0 mg.L-1 and IAA 0.1 mg.L-1 was shown to be more effective in Shoot regeneration than that complemented with BAP 2.0 mg.L-1 and IAA 0.1 mg.L-1. The frequency of shoot differentiation varied, ranging from 20 to 95%, but leaf segments provided better explants than the petioles for callus formation and shoot development. Guilio, one of cultivars of Cichorium intybus L. var. sativus was found to be the best genotype for shoot regeneration rate between 75 and 95%. It took only two weeks after the initial culture to develop the multiple shoots. Roots were differentiated when shoots were cultured on a half strength basal medium composition supplemented with NAA 0.1 mg.L-1. The sealing of the culture vessel was done with a cap having microspore membrane filters in its center to prevent vitrification by accelerating aeration.